Dietary supplement enhancing the muscular energy metabolism, comprising an alkanoyl carnitine and ribose

ABSTRACT

A method for treating myocardial or skeletal muscle anoxia which occurs in coronary or post-infarct disorders or during prolonged physical activity and muscle fatigue. This method comprises the administration of a combination composition comprising (a) an alkanoyl L-carnitine selected from the group consisting of isovaleryl L-carnitine, propionyl L-carnitine or the pharmacologically acceptable salts thereof or mixtures thereof; and (b) ribose or a phosphate derivative thereof.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 11/604,390filed on Nov. 27, 2006, which is a continuation-in-part of U.S.application Ser. No. 10/048,590 filed on Feb. 1, 2002, now abandoned,which is a 35 U.S.C. §371 national phase of PCT/IT01/00283 filed on Jun.1, 2001, which claims priority to and the benefit of Italian ApplicationNo. RM2000A000323 filed on Jun. 14, 2000, the contents of which areincorporated herein by reference.

BACKGROUND OF THE INVENTION

The present invention relates to a health food/dietary supplementcomprising as its characterising ingredients an alkanoyl L-carnitineselected from the group consisting of isovaleryl L-carnitine andpropionyl L-carnitine or their pharmacologically acceptable salts ormixtures of the same and a monosaccharide pentose, particularly riboseor its phosphorylated analogues.

It has been found that the above-mentioned composition is extremelyeffective in exerting a potent stimulation of muscular energymetabolism, and can thus be profitably used in the prevention ofmyocardial insufficiency and in post-infarct conditions, as well as inthe course of prolonged muscular effort during physical and sportingexercises, owing to the unexpected synergistic effect exerted by itscomponents.

Isovaleryl L-carnitine, a natural component of the pool of carnitines,presents specific activity at lysosomal level and on the cytosolicmovements of calcium. It is therefore capable of intervening inproteolytic processes such as occur during intense, prolonged effort andof protecting a number of organs, such as the liver, against the actionof toxic substances. Propionyl L-carnitine exerts an intense antioxidanteffect and is particularly effective in enhancing the peripheralcirculation and cardiac functional capacity.

Moreover, muscular carnitine transferase possesses a greater affinityfor propionyl L-carnitine than for L-carnitine, and consequentlypropionyl L-carnitine possesses a higher degree of specificity forcardiac and skeletal muscle. In addition, propionyl L-carnitinetransferase, transporting the propionyl group, increases the uptake ofthis component by the muscle cells, which may be of particularimportance for energy purposes, in that the propionate can be used bythe mitochondria as an anapleurotic substrate and provide energy in theabsence of oxygen.

Equally well known are the metabolic effects of ribose. Ribose is amonosaccharide pentose which is important in the body for the synthesisof nucleotides and other metabolic products. It is formed by conversionof glucose via the pentose phosphates. In the presence of a ribokinaseribose is phosphorylated to ribose-5-phosphate which, through theproduction of 5-phosphoribosyl-1-pyrophosphate (PRPP), can be used forthe synthesis of nucleotides necessary for the production of ATP. PRPP,in addition to intervening in the production of ATP, is also importantfor the synthesis of nucleotides such as adenine and hypoxanthine and ofribonucleotides and deoxyribonucleotides.

It has now been found surprisingly that a composition comprising acombination of the following as its characterizing components:

(a) an alkanoyl L-carnitine selected from the group comprisingisovaleryl L-carnitine, propionyl L-carnitine or their pharmacologicallyacceptable salts or mixtures of the same; and

(b) ribose or one of its phosphorylated derivatives thereof,

constitutes an effective health food/dietary supplement for theprevention of states of myocardial or skeletal muscle dysfunctionrelated to conditions of anoxia or insufficient energy supply asoccurring in coronary or post-infarct disorders or during prolongedphysical activity and muscle fatigue, owing to the potent and unexpectedsynergistic effect exerted by its components.

The weight-to-weight ratios of the above-mentioned components (a):(b)range from 1:1 to 1:10.

The dietary supplement according to the present invention mayadditionally contain

(c) a “carnitine” selected from the group comprising L-carnitine, acetylL-carnitine, butyryl L-carnitine and valeryl L-carnitine, or theirpharmacologically acceptable salts or mixtures of the same.

The weight-to-weight ratios of the above-mentioned components(a):(b):(c) range from 1:1:1 to 1:10:2.

The surprising synergistic effect achieved with the combination of“carnitines” (term denoting collectively both L-carnitine and thealkanoyl L-carnitines), particularly isovaleryl L-carnitine and/orpropionyl L-carnitine, and ribose, has been demonstrated by severalpharmacological tests (some of which are described here below) chosen insuch a way as to prove strongly predictive for the purposes of thepractical use of this composition in the preventive/nutritional/dieteticfield.

In particular, this unexpected synergistic effect on the increase inenergy capabilities at both cardiac and muscular level exerted by thecombination according to the present invention enables it to be used inthe prevention of both myocardial insufficiency and of muscle fatiguesuch as occur in cases of myocardial ischemia or in the course ofintense muscular effort due to prolonged physical exercise or sportingactivity.

DETAILED DESCRIPTION OF THE INVENTION Test of ATP Concentrations inHeart Subjected to Anoxia

In this test the technique adopted was the one using papillary muscle ofrabbit heart perfused and subjected to anoxia which, as is known, leadsto an impoverishment of its ATP energy reserves. With this test, the aimwas to observe whether or not preventive treatment with isovalerylL-carnitine, with propionyl L-carnitine, with a carnitine combination orwith ribose, or with a combination of these was capable of protectingcardiac muscle against the loss of ATP induced by anoxia.

In this test, a batch of New Zealand rabbits was used, subdivided intodifferent groups which were injected intravenously every day for threeconsecutive days with isovaleryl L-carnitine alone (100 mg/kg),propionyl L-carnitine alone (100 mg/kg) or a carnitine combinationconsisting of propionyl L-carnitine (25 mg/kg), acetyl L-carnitine (25mg/kg), L-carnitine (25 mg/kg), and isovaleryl L-carnitine (25 mg/kg) orwith ribose alone (100 mg/kg), or ribose combined with theabove-mentioned “carnitines”.

At the end of the third day of treatment, all the animals weresacrificed and their hearts excised. Sections of papillary musclemeasuring 1 mm in diameter and 4-5 mm in thickness were isolated fromthe excised hearts. The isolated papillary muscle was perfused in athermostatic bath with a saturated 100% O₂ solution.

The anoxic state was obtained by introducing 100% N₂ instead of O₂ intothe bath. For the measurement of the ATP concentrations in the papillarymuscle the method described by Strehler was adopted (Strehler B. L.Methods in Enzymology 111 N.Y. Acad. Press., 879, 1957).

The analysis was carried out on tissue samples maintained in conditionsof perfusion with oxygen for 90 minutes and after a period of anoxia ofthe same duration.

The results of this test, presented in Table 1, indicate that propionylL-carnitine, isovaleryl L-carnitine, the carnitine combination andribose are individually capable of partly protecting the ATP present inpapillary muscle against anoxia, but that it was only with thecombination of propionyl L-carnitine or isovaleryl L-carnitine plusribose or with the combination of the carnitine combination plus ribosethat complete protection against the anoxia-induced reduction in ATPcould be obtained, thus demonstrating the potent synergistic effectexerted by the components of the combination.

TABLE 1 Test of ATP concentrations in papillary muscle of heartsubjected to hypoxia ATP concentration (mol/g tissue) Treatment Beforehypoxia After hypoxia Controls 1.60 ± 0.55 0.41 ± 0.055 IsovalerylL-carnitine 1.50 ± 0.60 0.55 ± 0.65  Propionyl L-carnitine 1.64 ± 0.790.60 ± 0.040 Carnitine combination 1.55 ± 0.50 0.62 ± 0.060 Ribose 1.62± 0.39 0.55 ± 0.075 Isovaleryl L-carnitine + ribose 1.50 ± 0.25 1.15 ±0.055 Propionyl L-carnitine + ribose 1.61 ± 0.45 1.25 ± 0.35  Carnitinecombination + ribose 1.65 ± 0.60 1.16 ± 0.30 

Experimental Myocardial Anoxia Test

Adopting the technique described by Selych (Selych et al., Angiology,11, 398, 1960) and modified by Clark (Clark C., J. Pharmacol. Methods,3, 357, 1980), these tests were used to evaluate the protective activityof isovaleryl L-carnitine, propionyl L-carnitine, carnitine combination,ribose and various combinations of the same against ventriculararrhythmias induced by left coronary ligation in the rat.

Coronary occlusion and the resulting myocardial anoxia lead, after 5-8minutes, to the onset of arrhythmias. In these tests, ventricularectopic contractions were counted for a period of 30 minutes afterligation both in control rats and in rats that had received slowinjections into the left ventricle, 15 minutes before ligation, of asolution containing isovaleryl L-carnitine alone (100 mg/kg), propionylL-carnitine alone (100 mg/kg), or carnitine combination alone consistingof propionyl L-carnitine (25 mg/kg), acetyl L-carnitine (25 mg/kg) andisovaleryl L-carnitine (25 mg/kg) or ribose alone (100 mg/kg), or acombination of ribose plus isovaleryl L-carnitine or propionylL-carnitine or a combination of ribose plus carnitine combination at thedoses described above.

The results of this test (Table 2) indicate that, whereas isovalerylL-carnitine alone or propionyl L-carnitine alone or carnitinecombination alone or ribose alone produce only slight reductions in thenumber of ectopic contractions compared to controls, such contractionsare reduced almost to the extent of disappearing altogether when riboseis injected in combination with isovaleryl L-carnitine, or propionylL-carnitine, or carnitine combination, thus demonstrating the potent andunexpected synergistic effect exerted by the combination according tothe present invention.

TABLE 2 Test of arrhythmia induced by myocardial anoxia Start of N. ofectopic contractions arrhythmias during 30 minutes after Treatment after(mins) ligation Controls 5-7 989 ± 96 Isovaleryl L-carnitine 5-7 860 ±75 Propionyl L-carnitine 5-8 830 ± 86 Carnitine combination 5-8 810 ± 99Ribose 5-7  855 ± 110 Isovaleryl L-carnitine + ribose 6-7 270 ± 95Propionyl L-carnitine + ribose 6-8  230 ± 112 Carnitine combination +6-8 207 ± 93 ribose

Some non-limiting examples of compositions according to the presentinvention are given herein below:

Lozenges, capsules, tablets 1) Propionyl L-carnitine 500 mg Ribose 500mg 2) Isovaleryl L-carnitine 500 mg Ribose 500 mg 3) PropionylL-carnitine 125 mg Acetyl L-carnitine 125 mg L-carnitine 125 mgIsovaleryl L-carnitine 125 mg Ribose 500 mg Granulates or vials 4)Propionyl L-carnitine 1 g Ribose 1 g 5) Isovaleryl L-carnitine 1 gRibose 1 g 6) Propionyl L-carnitine 1 g Ribose 2.5 g 7) PropionylL-carnitine 250 mg Acetyl L-carnitine 250 mg Isovaleryl L-carnitine 250mg L-carnitine 250 mg Ribose 2.5 g 8) Propionyl L-carnitine 250 mgAcetyl L-carnitine 250 mg Isovaleryl L-carnitine 250 mg L-carnitine 250mg Ribose 2 g Ribonucleic acid 100 mg Deoxyribonucleic acid 100 mg 9)Propionyl L-carnitine 250 mg Acetyl L-carnitine 250 mg IsovalerylL-carnitine 250 mg L-carnitine 250 mg Ribose 2 g L-glutamine 100 mgL-alanine 100 mg L-arginine 100 mg L-glicine 100 mg L-histidine 100 mgL-isoleucine 100 mg L-phenylalanine 50 mg L-threonine 50 mg L-serine 100mg 10)  Propionyl L-carnitine 250 mg Acetyl L-carnitine 250 mgIsovaleryl L-carnitine 250 mg L-carnitine 250 mg Ribose 1 g Destrose 0.5g Fructose 0.5 g Maltose 0.5 g 11)  Propionyl L-carnitine 250 mg AcetylL-carnitine 250 mg Isovaleryl L-carnitine 250 mg L-carnitine 250 mgRibose 1 g Glucose-1,6-diphosphate 200 mg Fructose-1,6-diphosphate 200mg Galactose-1,6-phosphate 200 mg Glycerol-3-phosphate 200 mgPhosphenylpyruvate 100 mg Thiamine pyrophosphate 5 mgPyridoxal-5-phosphate 5 mg Magnesium stearate 2 mg 12)  PropionylL-carnitine 250 mg Acetil L-carnitine 250 mg Isovaleryl L-carnitine 250mg L-carnitine 250 mg Ribose 1 g Vit. A 1250 U.I. Vit. B₁ 0.5 mg Vit. B₆30 mg Vit. C 50 mg Vit. E 5 mg Nicotinammide 25 mg Vit. B₁₂ 100 mcg Vit.D 100 U.I. Pantothenic acid 30 mg Magnesium glycinate 5 mg Manganese 1mg L-Selenomethionine 50 mcg Molybdenum 10 mcg Zinc 1 mg

What is meant by a pharmacologically acceptable salt of the variouscarnitines mentioned in the present invention, is, in addition to therespective inner salts, any salt of these with an acid which does notgive rise to unwanted toxic or side effects. These acids are well knownto pharmacologists and to experts in pharmaceutical technology.

Non-limiting examples of such salts are the following: chloride;bromide; iodide; aspartate, acid aspartate; citrate, acid citrate;tartrate; phosphate, acid phosphate; fumarate, acid fumarate;glycerophosphate; glucose phosphate; lactate; maleate, acid maleate;mucate; orotate; oxalate, acid oxalate; sulphate, acid sulphate;trichloroacetate; trifluoroacetate and methane sulphonate.

Among these salts, isovaleryl L-carnitine acid fumarate (U.S. Pat. No.5,227,518) is particularly preferred.

A list of FDA-approved pharmacologically acceptable acids is given inInt. J. Pharm., 33, 1986, 201-217, the latter publication beingincorporated in the present specification by reference.

The supplement of the invention may further comprise vitamins,coenzymes, mineral substances, aminoacids and antioxidants. Thesupplement may be manufactured in the form of tablets, lozenges,capsules, pills, granulates, syrups, vials or drops.

While the invention has been described in connection with what ispresently considered to be the most practical and preferred embodiment,it is to be understood that the invention is not to be limited to thedisclosed embodiment, but on the contrary, is intended to cover variousmodifications and equivalents arrangements included within the spiritand scope of the appended claims.

The invention claimed is:
 1. A method for the treatment of states ofmyocardial or skeletal muscle anoxia occurring in coronary orpost-infarct disorders or during prolonged physical activity and musclefatigue, which comprises administering to an individual in need thereofa combination composition consisting of the following ingredients: (a)an alkanoyl L-carnitine selected from the group consisting of isovalerylL-carnitine, propionyl L-carnitine or the pharmacologically acceptablesalts thereof or mixtures thereof; and (b) ribose.
 2. The method ofclaim 1, wherein the weight ratio of ingredients (a):(b) ranges from 1:1to 1:10.
 3. A method for the treatment of states of myocardial orskeletal muscle anoxia occurring in coronary or post-infarct disordersor during prolonged physical activity and muscle fatigue, whichcomprises administering to an individual in need thereof a combinationcomposition consisting of the following ingredients: (a) an alkanoylL-carnitine selected from the group consisting of isovalerylL-carnitine, propionyl L-carnitine or the pharmacologically acceptablesalts thereof or mixtures thereof; (b) ribose; and (c) vitamins, sugars,coenzymes, mineral substances, amino acids, peptides and antioxidants.4. The combination composition of claim 1 wherein the pharmacologicallyacceptable salt is selected from the group consisting of chloride,bromide, iodide, aspartate, acid aspartate, citrate, acid citrate,tartrate, phosphate, acid phosphate, fumarate, acid fumarate,glycerophosphate, glucose phosphate, lactate, maleate, acid maleate,mucate, orotate, oxalate, acid oxalate, sulfate, acid sulfate,trichloracetate, trifluoroacetate and methane sulfonate.
 5. The methodof claim 1, wherein said combination composition is in form of tablets,capsules, lozenges, pills, granules, creams, syrups or drops.
 6. Amethod for the treatment of states of myocardial or skeletal muscledysfunction related to conditions of anoxia or insufficient energysupply as occurring in coronary or post-infarct disorders or duringprolonged physical activity and muscle fatigue, which comprisesadministering to an individual in need thereof a combination compositionconsisting of the following ingredients: (a) an alkanoyl L-carnitineselected from the group consisting of isovaleryl L-carnitine, propionylL-carnitine or the pharmacologically acceptable salts thereof or mixturethereof, and (b) ribose.
 7. The method of claim 6, wherein the weightratio of ingredients (a):(b) ranges from 1:1 to 1:10.
 8. A method forthe treatment of states of myocardial or skeletal muscle dysfunctionrelated to conditions of anoxia or insufficient energy supply asoccurring in coronary or post-infarct disorders or during prolongedphysical activity and muscle fatigue, which comprises administering toan individual in need thereof a combination composition consisting ofthe following ingredients: (a) an alkanoyl L-carnitine selected from thegroup consisting of isovaleryl L-carnitine, propionyl L-carnitine or thepharmacologically acceptable salts thereof or mixture thereof; (b)ribose; and (c) vitamins, sugars, coenzymes, mineral substances, aminoacids, peptides and antioxidants.
 9. The combination composition ofclaim 6 wherein the pharmacologically acceptable salt is selected fromthe group consisting of chloride, bromide, iodide, aspartate, acidaspartate, citrate, acid citrate, tartrate, phosphate, acid phosphate,fumarate, acid fumarate, glycerophosphate, glucose phosphate, lactate,maleate, acid maleate, mucate, orotate, oxalate, acid oxalate, sulfate,acid sulfate, trichloracetate, trifluoroacetate and methane sulfonate.10. The method of claim 6, wherein said combination composition is inform of tablets, capsules, lozenges, pills, granules, creams, syrups ordrops.
 11. A method for reducing arrhythmia induced by myocardialanoxia, said method comprising administering to an individual in needthereof a combination composition consisting the following ingredients:(a) isovaleryl L-carnitine and propionyl L-carnitine or thepharmacologically acceptable salt thereof; and (b) ribose, wherein theweight ratio of ingredients (a):(b) ranges from 1:1 to 1:10; and (c)vitamins, sugars, coenzymes, mineral substrates, amino acids, peptidesand antioxidants.
 12. The method of claim 11, wherein saidpharmacologically acceptable salt is selected from the group consistingof chloride, bromide, iodide, aspartate, acid aspartate, citrate, acidcitrate, tartrate, phosphate, acid phosphate, fumarate, acid fumarate,glycerophosphate, glucose phosphate, lactate, maleate, acid maleate,mucate, ororate, oxalate, acid oxalate, sulphate, acid sulphate,trichloroacetate, trifluoroacetate and methane sulfonate.
 13. The methodof claim 11, wherein said combination composition is in form of tablets,capsules, lozenges, pills, granules, creams, syrups or drops.